冠状动脉粥样硬化模型猪胆固醇代谢异常对Aβ的影响及复方干预研究

发布时间：2018-06-04 04:44

本文选题：β淀粉样蛋白 + 阿尔茨海默病　；参考：《辽宁中医药大学》2015年博士论文

【摘要】：目的:通过高脂饮食建立高脂致Aβ增高小型猪动物模型,观察高胆固醇血症胆固醇代谢异常状态对脑内β淀粉样蛋白生成的影响,探讨胆固醇代谢异常状态下脑胆固醇代谢紊乱相关因素与AD早期病理改变β淀粉样蛋白生成的关系;运用具有不同功效的中药复方干预此动物模型,并对其作用机制进行探讨,为在早期阶段通过减少AD典型病理特征Aβ生成及沉积从而防治AD提供实验依据。材料与方法:广西巴马小型猪55只,月龄6-8个月,体重(29.88±2.27)kg,雄性,肠道菌群正常,体健,由泰州泰和生物科技有限公司提供。伦理符合中华人民共和国卫生部动物实验管理条例(No.55,2001)和辽宁中医药大学实验动物管理条例规定。实验室温度为(20.80±2.97)℃,湿度(47.30±15.31)%,实验动物均为单笼饲养。适应性饲养1周后,采用随机数字表法将55只小型猪分为两组:正常组5只,手术组50只。正常组给予基础饮食,手术组(第2周进行冠脉球囊挤压伴拉伤,术前禁食8h,禁水2小时),术后继续予以高热量高脂适量胆盐喂饲(防止小猪连续吃大量油腻食物出现厌食,加胆盐是防止消化不良)。日进食量为上周所称体重的3%,自由饮水。术中有8只小型猪死亡;应用随机数字表将存活的42只小型猪重新进行随机分组:模型组(n=7)、健脾祛痰化瘀组(n=7)、健脾祛痰组(n=7)、健脾组(n=7)、祛痰化瘀组(n=7)、西药组(n=7)至此,健脾组加入健脾方,健脾祛痰组加入健脾祛痰方,祛痰化瘀组加入祛痰化瘀方,健脾祛痰化瘀复方组加入健脾祛痰化瘀方,加入相应剂量的中药复方按照体重对应标准,饲喂至24周,指标检测如下:1.实验期间每4周称量小型猪的体重,观察小型猪体重变化情况。2.实验第0周及实验开始后第4周,第12周,第24周禁食16h后耳缘静脉抽血3ml,全自动生化分析仪检测血脂(TC、TG、LDL、HDL)水平,观察不同时间点血脂四项的变化情况。3.免疫组织化学染色法检测Aβ阳性细胞在大脑组织的分布。4.免疫组织化学法观察Aβ在大脑组织的含量及水平。5.免疫组织化学法检测Aβ代谢相关因子BACE1及脑胆固醇代谢关键因子HMG-Co AR、CYP46a、ABCA1在脑内的含量及在神经元细胞内的表达。6.免疫蛋白印迹法(Western Blot)检测Aβ代谢相关因子BACE1及脑胆固醇代谢关键因子HMG-Co AR、CYP46a、Apo E、ABCA1的蛋白表达。本实验所有数据结果使用SPSS17.0软件进行统计分析,结果用平均值加减标准差(—X±S)表示,组与组之间比较采用单因素方差分析。(P0.05说明有显著性差异,P0.01说明有非常显著性差异。)结果:1.正常组与模型组小型猪的体重增长变化显示:模型组与正常组在第0周体重增长并无明显差异,两组巴马小型猪体重增长从第4周起开始有显著性差异(P0.05);2.正常小猪在成长过程中,血脂的变化不明显;而模型组巴马小型猪血脂四项变化显著,结果显示:高脂饮食1周,可使小型猪甘油三酯升高2.86倍,使总胆固醇升高3.84倍,使低密度脂蛋白升高7.40倍,使高密度脂蛋白升高2.56倍;随着高脂饮食时间延长,小型猪甘油三酯(TG)、总胆固醇(TC)、低密度脂蛋白(LDL)持续升高,第24周趋于稳定,而高密度脂蛋白随高脂饮食时间增加呈降低趋势;3.免疫组织化学法显示高脂饮食造成的高血胆固醇水平增加了脑Aβ含量,模型组巴马小型猪脑Aβ含量显著高于正常组(0.26±0.03 pg/mg protein vs 0.1110±0.01 pg/mg protein,p0.01)。免疫组织化学法检测平均光密度值(AOD)正常组与模型组相比较,有统计学意义。(P0.01)。免疫组织化学染色显示与正常组相比较,模型组脑组织神经元细胞结构呈棕黄色颗粒聚集,表示脑组织内Aβ阳性细胞表达增多;4.健脾祛痰组调节TC、HDL、LDL的能力强于其余两组,健脾祛痰化瘀组调节TG的能力强于其余两组;5.与模型组比较,健脾组、健脾祛痰组、祛痰化瘀组、健脾祛痰化瘀组均能明显降低脑内Aβ含量(P0.01);健脾祛痰化瘀组较健脾组、健脾祛痰组、祛痰化瘀组减少Aβ含量,且有统计学意义(P0.01)健脾组、健脾祛痰组、祛痰化瘀组之间比较,无统计学差异(P0.05);6.经健脾祛痰化瘀法干预后,健脾组、健脾祛痰组、祛痰化瘀组、健脾祛痰化瘀组脑内BACE1蛋白表达明显降低;证明血胆固醇降低引起的脑Aβ降低与Aβ生成相关因子的蛋白表达的改变有关;7.免疫组织化学法检测ABCA1实验结果显示,与正常组相比较,模型组及中药复方各组脑组织ABCA1含量增加,差异有统计学意义(P0.01);与模型组相比较,健脾组、健脾祛痰组、祛痰化瘀组、健脾祛痰化瘀组ABCA1含量升高,差异有统计学意义(P0.01);与健脾祛痰化瘀组相比,健脾组、健脾祛痰组、祛痰化瘀组ABCA1含量降低,差异有统计学意义(P0.01);健脾组、健脾祛痰组、祛痰化瘀组组间比较差异无统计学意义(P0.01)。免疫组化染色显示模型组阳性细胞表达增多,而药物组阳性细胞表达减少,与模型组相比,差异有统计学意义(P0.01);8.免疫组织化学法检测HMG-Co AR结果显示,与正常组相比,模型组及中药复方各组脑组织HMG-Co A含量增加,且有显著性差异(P0.01);与模型组相比较,健脾组、健脾祛痰组、祛痰化瘀组及健脾祛痰化瘀组HMG-Co AR含量增加,差异有统计学意义(P0.01);与健脾祛痰化瘀组相比,健脾组,健脾祛痰组、祛痰化瘀组HMG-Co AR含量减少,差异有统计学意义(P0.01);健脾组,健脾祛痰组,祛痰化瘀组之间比较,HMG-Co AR差异无统计学意义(P0.01),免疫组化染色显示模型组阳性细胞表达增多,而药物组阳性细胞表达减少,与模型组相比,差异有统计学意义;9.免疫组织化学法检测CYP46a结果显示,与正常组相比较,模型组、复方各组脑组织CYP46a含量增加,且有显著性差异(P0.01);与模型组相比较,复方组脑组织内CYP46a含量增加,差异有统计学意义(P0.01);与健脾祛痰化瘀组相比,健脾组、健脾祛痰组、祛痰化瘀组脑组织CYP46a含量减少,差异有统计学意义(P0.01);与健脾组相比,健脾祛痰组、祛痰化瘀组脑组织CYP46a含量增加,且差异有统计学意义(P0.01);与健脾祛痰组相比,祛痰化瘀组CYP46减少,差异无统计学意义。(P0.05)。免疫组化染色显示模型组阳性细胞表达增多,而药物组阳性细胞表达减少,与模型组相比,差异有统计学意义(P0.01);10.Western Blot结果显示,与正常组相比较,模型组、中药复方组脑组织HMG-Co AR蛋白表达升高,且差异有统计学意义(P0.01);与模型组相比较,复方组HMGR蛋白表达升高,且差异有统计学意义(P0.01);与健脾祛痰化瘀组相比,健脾组、健脾祛痰组、祛痰化瘀组脑组织HMG-Co AR蛋白表达降低,差异有统计学意义(P0.01)与健脾组相比,健脾祛痰组、祛痰化瘀组HMG-Co AR蛋白表达升高,差异有统计学意义(P0.05);健脾祛痰组、祛痰化瘀组相比,HMGR蛋白表达差异无统计学意义(P0.05);11.Western Blot结果显示,与正常组相比,模型组、复方组脑组织CYP46a蛋白表达升高,差异有统计学意义(P0.01);与模型组相比较,复方组脑组织内CYP46a1蛋白表达均有升高,差异有统计学意义(P0.01);与健脾祛痰化瘀相比,祛痰化瘀组、健脾祛痰组及健脾组脑组织CYP46a蛋白表达降低,差异有统计学意义;健脾组、健脾祛痰组、祛痰化瘀组脑组织CYP46a1蛋白表达无显著性差异。(P0.05);12.Western Blot结果显示,与正常组相比较,模型组、复方组脑组织Apo E蛋白表达升高,且有显著性差异(P0.01);与模型组相比,复方组脑组织内Apo E蛋白表达降低,差异有统计学意义(P0.01);与健脾祛痰化瘀组相比较,祛痰化瘀组、健脾祛痰组及健脾组脑组织Apo E蛋白表达升高,有显著性差异(P0.01);健脾组、健脾祛痰组、祛痰化瘀组脑组织蛋白表达差异无统计学意义。(P0.05);13.Western Blot结果显示,与正常组相比,模型组、复方组脑组织ABCA1蛋白表达升高,且有显著性差异(P0.01);与模型组相比较,复方组对脑组织内ABCA1蛋白表达升高,差异有统计学意义(P0.01);与健脾祛痰化瘀组相比,健脾祛痰组、祛痰化瘀组及健脾组脑组织ABCA1蛋白表达降低,差异有统计学意义(P0.01);健脾组、健脾祛痰组、祛痰化瘀组之间相比,脑组织ABCA1蛋白表达无显著性差异。(P0.05)。结论:1.高脂饮食可影响血脂四项水平,高脂饮食1周可造成冠状动脉粥样硬化模型猪血脂异常,随高脂饮食时间延长,血脂四项中甘油三酯(TG)、总胆固醇(TC)、低密度脂蛋白(LDL)的浓度升高,高密度脂蛋白(HDL)浓度降低;2.高脂饮食增加了外周血血脂浓度,外周血脂水平可影响巴马小型猪脑内Aβ的水平的改变,使脑内Aβ含量增加,高脂饮食24周可建立高脂致Aβ增高动物模型;3.健脾方、健脾祛痰方、祛痰化瘀方、健脾祛痰化瘀方可调节巴马小型猪模型的血脂水平,健脾祛痰化瘀方优于健脾、健脾祛痰及祛痰化瘀方;4.健脾方、健脾祛痰方、祛痰化瘀方、健脾祛痰化瘀方可减少Aβ在脑内的沉积,减少Aβ的表达;5.健脾方、健脾祛痰方、祛痰化瘀方、健脾祛痰化瘀方减少Aβ含量和生成的可能机制之一是通过降低Aβ代谢关键因子BACE1蛋白表达来实现的;6.脑胆固醇代谢异常能够影响Aβ生成与代谢;7.健脾方、健脾祛痰方、祛痰化瘀方、健脾祛痰化瘀方可使脑中胆固醇合成及转出的关键酶Apo E蛋白下调,使HMGR、ABCA1、和CYP46的蛋白表达上调,其中健脾祛痰化瘀方作用优于健脾、健脾祛痰、祛痰化瘀、健脾祛痰化瘀方。8.健脾方、健脾祛痰方、祛痰化瘀方、健脾祛痰化瘀方干预高脂所致Aβ增高动物模型的可能机制是通过下调Apo E蛋白表达,上调HMGR、ABCA1、和CYP46的蛋白表达来发挥作用。9.健脾方、健脾祛痰方、祛痰化瘀方、健脾祛痰化瘀方可能是通过调节巴马小型猪的外周血脂水平、调节Aβ生成相关分泌酶β分泌酶的活性及BACE1蛋白表达、下调Apo E蛋白表达,上调CYP46a HMG-Co AR、ABCA1等蛋白的表达来干预阿尔茨海默病早期β淀粉样蛋白形成的病理过程,从而在早期阻抑或延缓AD的发生发展。10.健脾方、健脾祛痰方、祛痰化瘀方、健脾祛痰化瘀方在调脂、调节Aβ生成及改善脑胆固醇代谢方面效果显著,健脾祛痰化瘀方作用大于健脾祛痰方、祛痰化瘀方大于健脾方。说明健脾祛痰化瘀方配伍精良,健脾、祛瘀、化痰三者均不可少。
[Abstract]:Objective: to establish an animal model of high fat induced A beta hyperlipidemia pig by high fat diet, to observe the effect of abnormal cholesterol metabolism in hypercholesterolemia on the formation of beta amyloid in the brain, and to explore the relationship between the related factors of cholesterol metabolism in the abnormal state of cholesterol metabolism and the early pathological changes of beta amyloid protein in AD. In order to provide experimental basis for the prevention and treatment of AD by reducing the typical pathological features of AD, A beta generation and deposition in the early stage, 55 Chinese Bama miniature pigs, 6-8 months of age, weight (29.88 + 2.27) kg, male and normal intestinal flora, were provided for the prevention and treatment of AD in the early stage by reducing the typical pathological characteristics of AD. Body health, provided by Taizhou Taihe biological science and Technology Co., Ltd., the ethics conforms to the regulations of the animal experiment management regulation (No.552001) of the Ministry of health of People's Republic of China and the regulations of the experimental animal management of Liaoning University of Traditional Chinese Medicine. The laboratory temperature is (20.80 + 2.97) and humidity (47.30 + 15.31)%, the experimental animals are kept in single cage. After 1 weeks of adaptive breeding, 55 small pigs were divided into two groups by random digital table: 5 in the normal group and 50 in the operation group. The normal group was given the basic diet, the operation group (second weeks of coronary balloon extrusion accompanied by pull, the preoperative fasting 8h, the water prohibition 2 hours), and the postoperative continuous high calorie, high fat and moderate salt feeding (preventing the piglets to eat a large amount of greasy food in an anorexia. Bile salt is to prevent dyspepsia). Intake of daily intake is 3% of weight of last week's weight, free drinking water. 8 small pigs died in the operation; 42 small pigs survived were randomly divided into model group (n=7), spleen eliminating phlegm and stasis group (n=7), spleen eliminating phlegm group (n=7), spleen invigorating group (n=7), phlegm removing stasis group (n=7), western medicine group (n=7) The Jianpi group joined Jianpi prescription, Jianpi dispelling phlegm group joined Jianpi dispelling phlegm prescription, dispelling phlegm and removing stasis group joined the prescription of eliminating phlegm and removing stasis, the compound group of Jianpi dispelling phlegm and removing stasis was added to Jianpi dispelling phlegm and removing stasis prescription, and the corresponding dosage of Chinese medicine compound was added to the body weight corresponding standard for 24 weeks, and the indexes were measured as follows: the weight of small pigs was weighed every 4 weeks during the 1. experiment. The body weight change of small pigs was examined by.2. zeroth weeks and fourth weeks, Twelfth weeks and twenty-fourth weeks after the experiment. 3ml was extracted after 16h, and the level of blood lipid (TC, TG, LDL, HDL) was detected by automatic biochemical analyzer, and the changes of blood lipid four items at different time points were observed..3. free histochemical staining was used to detect the expression of A beta positive cells in the brain tissue. The content and level of A beta in brain tissue were observed by distributed.4. immunohistochemical method and.5. immunohistochemical staining was used to detect A beta metabolism related factor BACE1 and the key factor of brain cholesterol metabolism HMG-Co AR, CYP46a, ABCA1 in the brain and the expression of.6. immunoblotting (Western Blot) in the neuron cells. BACE1 and the key factors of brain cholesterol metabolism HMG-Co AR, CYP46a, Apo E, ABCA1 protein expression. All data results in this experiment were statistically analyzed using SPSS17.0 software, and the results were added and subtracting the standard deviation (- X + S) with the average value (- X + S), and the group and the group were compared with the single factor variance analysis. (P0.05 shows a significant difference, P0.01 instructions have a very significant difference. Results: the body weight growth of 1. normal groups and model group small pigs showed that there was no significant difference between the model group and the normal group at zeroth weeks. The weight growth of the two groups of Bama miniature pigs began to have significant difference from fourth weeks (P0.05); 2. normal piglets had no significant changes in blood lipid during the growth process; The four changes of blood lipid were significant. The results showed that high fat diet for 1 weeks could increase triglyceride of small pigs by 2.86 times, increase total cholesterol by 3.84 times, increase low density lipoprotein by 7.40 times, make high density lipoprotein (HDL) increase 2.56 times. With the prolongation of high fat diet, small pig triglyceride (TG), total cholesterol (TC), low density lipoprotein (LDL) (LDL) continued to rise, and the twenty-fourth weeks tended to stabilize, while high density lipoprotein decreased with the increase of high fat diet time; 3. the high blood cholesterol level caused by high fat diet increased the content of brain A beta in the high fat diet, and the A beta content in the brain of the model group was significantly higher than that of the normal group (0.26 + 0.03 pg/mg protein vs 0.1110 + 0.01 pg/m G protein, P0.01). Compared with the model group, the average light density (AOD) in the normal group was compared with the model group. (P0.01). (P0.01). Immunohistochemical staining showed that the structure of neurons in the brain tissue of the model group was brown yellow particles gathering, indicating that the expression of A beta positive cells in the brain tissue increased; 4. The ability of regulating TC, HDL, LDL in the phlegm group is stronger than the other two groups, and the ability of regulating TG in the invigorating spleen and removing phlegm and removing stasis group is stronger than the other two groups. 5. compared with the model group, the group of invigorating the spleen, invigorating the spleen and removing phlegm, removing phlegm and removing stasis, and eliminating phlegm and removing stasis group can obviously reduce the content of A beta in the brain (P0.01); the group of Jianpi dispelling phlegm and removing stasis is more than the invigorating spleen group, invigorating the spleen and expectorant group and removing phlegm and removing stasis group. Reduce the content of A beta, and have statistical significance (P0.01) in the invigorating spleen group, invigorating the spleen and eliminating phlegm group, eliminating phlegm and removing stasis group, there is no statistical difference (P0.05); 6. after strengthening the spleen and removing phlegm and removing stasis method, the expression of BACE1 protein in the brain of Jianpi group, Jianpi dispelling phlegm group, dispelling phlegm and removing stasis in the brain of Jianpi dispelling phlegm and removing stasis is obviously reduced, and the brain A beta caused by the decrease of blood cholesterol is proved to be the brain A beta The changes in the protein expression of A beta generation related factors were reduced; 7. the results of ABCA1 test by immunohistochemistry showed that the content of ABCA1 in the brain tissue of the model group and the traditional Chinese medicine compound was increased compared with the normal group, and the difference was statistically significant (P0.01); compared with the model group, the spleen strengthening group, the invigorating spleen and eliminating phlegm group, eliminating phlegm and removing stasis group and invigorating the spleen and removing the spleen were compared with the model group. The content of ABCA1 in the phlegm and removing stasis group was higher, the difference was statistically significant (P0.01). Compared with the invigorating spleen and eliminating phlegm and removing stasis group, the content of ABCA1 in the Jianpi group, the invigorating spleen and eliminating phlegm group and the dispelling phlegm and removing stasis group were statistically significant (P0.01), and there was no significant difference between the Jianpi group, the invigorating spleen and eliminating phlegm group and the expectorant and removing stasis group (P0.01). Immunohistochemical staining display model was not significant. The expression of positive cells in the group increased and the expression of positive cells in the drug group decreased. Compared with the model group, the difference was statistically significant (P0.01). The results of HMG-Co AR by 8. immunohistochemical staining showed that the content of HMG-Co A in the brain tissues of the model group and the traditional Chinese medicine group was significantly higher than that in the normal group, and there were significant differences (P0.01), compared with the model group, The content of HMG-Co AR in Jianpi group, invigorating spleen and eliminating phlegm group, eliminating phlegm and removing stasis group and promoting phlegm and removing stasis group increased with statistical significance (P0.01). Compared with the group of Jianpi dispelling phlegm and removing stasis, the content of HMG-Co AR in Jianpi group, Jianpi dispelling phlegm group and dispelling phlegm and removing stasis group was statistically significant (P0.01), and the comparison between Jianpi group, Jianpi dispelling phlegm group and eliminating phlegm and removing stasis group was compared. The difference of HMG-Co AR was not statistically significant (P0.01). Immunohistochemical staining showed that the expression of positive cells in the model group increased, but the expression of positive cells in the drug group decreased, and the difference was statistically significant compared with the model group. 9. the results of CYP46a detection by immunohistochemistry showed that the content of CYP46a in the brain tissue of the model group and the compound group was increased compared with the normal group. And there was significant difference (P0.01). Compared with the model group, the content of CYP46a in the brain tissue of the compound group increased, and the difference was statistically significant (P0.01). Compared with the invigorating spleen and eliminating phlegm and removing stasis group, the content of CYP46a in the brain tissue of the invigorating spleen group, the invigorating spleen and eliminating phlegm group and the dispelling phlegm and removing stasis group was statistically significant (P0.01); compared with the Jianpi group, the spleen eliminating phlegm group and expectorant were compared with the invigorating spleen group. The content of CYP46a in the brain tissue of the Huayu group increased and the difference was statistically significant (P0.01). Compared with the invigorating spleen and eliminating phlegm group, the difference was not statistically significant (P0.05). (P0.05). Immunohistochemical staining showed that the expression of positive cells in the model group increased and the positive cell expression of the drug group decreased, compared with the model group, the difference was statistically significant (P0. 01); the results of 10.Western Blot showed that the expression of HMG-Co AR protein in the brain tissue of the model group and the compound group was higher than that of the normal group, and the difference was statistically significant (P0.01). Compared with the model group, the expression of HMGR protein in the compound group was higher, and the difference was statistically significant (P0.01). Compared with the group of invigorating the spleen and removing phlegm and removing stasis, the spleen and the spleen and eliminating phlegm group were compared. The expression of HMG-Co AR protein in the brain tissue of the dispelling phlegm and removing stasis group decreased, and the difference was statistically significant (P0.01), compared with the Jianpi group, the expression of HMG-Co AR protein in the group of invigorating the spleen and removing phlegm and removing stasis was statistically significant (P0.05). There was no significant difference in the expression of HMGR protein in the group of invigorating the spleen and eliminating phlegm and removing phlegm and removing stasis (P0.05) and 11.Western Blot junction. Compared with the normal group, the expression of CYP46a protein in the brain tissue of the model group and the compound group increased, and the difference was statistically significant (P0.01). Compared with the model group, the expression of CYP46a1 protein in the brain tissue of the compound group increased, and the difference was statistically significant (P0.01). Compared with the invigorating spleen and eliminating phlegm and removing stasis, the group of dispelling phlegm and removing stasis, strengthening the spleen and eliminating phlegm and invigorating the spleen group brain group The expression of CYP46a protein decreased, and the difference was statistically significant. There was no significant difference in the expression of CYP46a1 protein in the brain tissue of the Jianpi group, the invigorating spleen and eliminating phlegm group and the removing phlegm and removing stasis group. (P0.05); the results of 12.Western Blot showed that the expression of egg white in the model group and the compound group brain tissue was higher than that of the normal group, and there was a significant difference (P0.01) in the model group (P0.01). Compared with the model group, the expression of the protein expression was significantly higher than that of the normal group (P0.01). The expression of Apo E protein in the brain tissue of the compound group decreased (P0.01), and the expression of Apo E protein in the brain tissue of the anti phlegm and removing stasis group, the invigorating spleen and eliminating phlegm group and the invigorating spleen group increased significantly (P0.01), and there was no significant difference in the protein expression of the brain tissue in the Jianpi group, the invigorating spleen and eliminating phlegm group and the dispelling phlegm and removing stasis group. (P0.05); the results of 13.Western Blot showed that the expression of ABCA1 protein in the brain tissue of the model group and the compound group was higher than that in the normal group, and there was a significant difference (P0.01). Compared with the model group, the expression of ABCA1 protein in the brain tissue was higher, the difference was statistically significant (P0.01). Compared with the group of invigorating the spleen and removing phlegm and removing stasis, the group of invigorating the spleen and removing phlegm and removing phlegm and removing stasis The expression of ABCA1 protein in the brain tissue of the group and the invigorating spleen group decreased, and the difference was statistically significant (P0.01). There was no significant difference in the expression of ABCA1 protein in the brain tissue between the Jianpi group, the invigorating spleen and eliminating phlegm group and the dispelling phlegm and removing stasis group. (P0.05) 1. high fat diet could affect the level of blood lipid, and the high fat diet could cause coronary atherosclerosis model pig blood for 1 weeks. Lipid abnormality, with the prolongation of high fat diet time, the concentration of triglyceride (TG), total cholesterol (TC), low density lipoprotein (LDL) and high density lipoprotein (HDL) decreased in four lipids, and 2. high fat diet increased the concentration of blood lipid in peripheral blood, and the level of peripheral blood lipid could affect the level of A beta in the brain of Bama small pig, and increase the content of A beta in the brain. Plus, high fat diet for 24 weeks can establish high fat induced A beta animal model; 3. Jianpi Recipe, invigorating the spleen and eliminating phlegm prescription, dispelling phlegm and removing stasis prescription, invigorating the spleen and Removing Blood Stasis Recipe can regulate the blood lipid level of Bama miniature pig model, strengthening spleen and removing phlegm and removing stasis prescription is superior to the spleen, invigorating the spleen and eliminating phlegm and removing phlegm and removing stasis; 4. invigorating spleen, eliminating phlegm and removing phlegm, eliminating phlegm and removing stasis, invigorating the spleen and removing phlegm and removing stasis It can reduce the deposition of A beta in the brain and reduce the expression of A beta; 5. the prescription of Jianpi Fang, Jianpi Dispelling Phlegm Recipe, dispelling phlegm and removing stasis, reducing the content and formation of A beta by eliminating phlegm and removing stasis is realized by reducing the expression of BACE1 protein, the key factor of A beta metabolism; the 6. brain cholesterol metabolism abnormality can affect the formation and metabolism of A beta; 7. spleen invigorating prescription, The recipe for invigorating the spleen and removing phlegm, eliminating phlegm and removing stasis, can reduce the protein expression of the key enzyme of cholesterol synthesis and transformation in the brain, and make the expression of the protein of HMGR, ABCA1, and CYP46 up regulation. The effect of strengthening spleen and removing phlegm and removing stasis is superior to the spleen strengthening, invigorating the spleen and eliminating phlegm, removing phlegm and removing stasis, strengthening spleen and removing phlegm and removing blood stasis prescription, invigorating spleen and eliminating phlegm, dispelling phlegm and removing stasis The possible mechanism of invigorating spleen and removing phlegm and removing stasis to interfere with high fat A beta animal model is to regulate the expression of Apo E protein, increase the expression of HMGR, ABCA1, and CYP46 to play the role of.9. Jianpi Recipe, invigorate the spleen and dispelling phlegm prescription, dispelling phlegm and removing stasis prescription, the prescription for regulating spleen and removing phlegm and removing stasis may be regulated by the peripheral blood lipid level of Bama miniature pig, regulating A Beta production of secretory enzyme beta secretase activity and BACE1 protein expression, downregulation of Apo E protein expression, up regulation of CYP46a HMG-Co AR, ABCA1 and other proteins
【学位授予单位】：辽宁中医药大学
【学位级别】：博士
【学位授予年份】：2015
【分类号】：R-332;R543.3

【参考文献】